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黏液型和非黏液型铜绿假单胞菌对仓鼠气管上皮细胞黏附的定量分析

Quantitation of adherence of mucoid and nonmucoid Pseudomonas aeruginosa to hamster tracheal epithelium.

作者信息

Marcus H, Baker N R

出版信息

Infect Immun. 1985 Mar;47(3):723-9. doi: 10.1128/iai.47.3.723-729.1985.

Abstract

Adherence of mucoid and nonmucoid isolates of Pseudomonas aeruginosa to tracheal epithelium was quantitated by using hamster tracheas mounted in a perfusion chamber. The strains of P. aeruginosa used were clinical isolates from cystic fibrosis patients and a series of laboratory strains. Aseptically excised hamster tracheas were mounted in perfusion chambers and embedded in minimal essential medium containing 1.5% agarose. The tracheas were infected with various numbers of bacteria for various periods, rinsed, homogenized, and plated on Trypticase soy agar. A 4-mm segment from each trachea was prepared for quantitation, and the other segment was prepared for examination by scanning electron microscopy. Adherence increased with time and with increasing concentrations of inoculum. Standard conditions of inoculation were set at an inoculum of 10(7) CFU/ml and a 2-h incubation. Under these conditions, the mucoid organisms adhered to the ciliated epithelium 10- to 100-fold better than did the nonmucoid organisms. Adherence of the mucoid isolates did not appear to be pilus mediated and did not involve hydrophobic interactions. The mucoid P. aeruginosa isolates could be seen adhering to the epithelium in the form of microcolonies embedded in an extracellular matrix which attaches the organisms to the cilia and to each other. The adherence may be involved in the establishment of infection of the lungs of these patients and in the inability to clear the organisms from the lungs. The model will be useful in determining the mechanism of adherence of the bacteria to the ciliated epithelium of the respiratory tract.

摘要

通过使用安装在灌注室中的仓鼠气管,对铜绿假单胞菌的黏液型和非黏液型分离株与气管上皮的黏附进行了定量分析。所用的铜绿假单胞菌菌株为来自囊性纤维化患者的临床分离株和一系列实验室菌株。无菌切除的仓鼠气管被安装在灌注室中,并包埋在含有1.5%琼脂糖的最低必需培养基中。气管在不同时间段感染不同数量的细菌,冲洗、匀浆后接种于胰蛋白酶大豆琼脂平板上。从每个气管切取4毫米长的片段用于定量分析,另一段用于扫描电子显微镜检查。黏附随着时间和接种物浓度的增加而增加。接种的标准条件设定为接种物浓度为10(7) CFU/ml,孵育2小时。在这些条件下,黏液型菌株比非黏液型菌株与纤毛上皮的黏附能力强10至100倍。黏液型分离株的黏附似乎不是由菌毛介导的,也不涉及疏水相互作用。可以看到黏液型铜绿假单胞菌分离株以微菌落的形式黏附在上皮细胞上,这些微菌落嵌入细胞外基质中,将细菌与纤毛以及彼此连接起来。这种黏附可能与这些患者肺部感染的发生以及无法从肺部清除细菌有关。该模型将有助于确定细菌与呼吸道纤毛上皮黏附的机制。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cdf6/261370/06dcf3139327/iai00120-0154-a.jpg

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